FIGURE 4.

Berberine inhibits the migration of TNBC cells in a LH2-dependent manner. (A–B). The secretion of IL-6 was detected by using the IL-6 Elisa kit. MDA-MB-231 cells were cultured with or without berberine (1.25, 2.5 and 5 μM) for 48 h, then the supernatant medium was collected for the detection of IL-6 levels. (B) Recombination human IL-6 protein (5 ng/ml) was used to stimulate MDA-MB-231 cells for 6 h, and berberine (5 μM) was used to treat MDA-MB-231 cells for 48 h, following which the cells were collected for transwell assay for 12 h. (C–D) LH2 mRNA and protein expression in the MDA-MB-231 and BT-549 cells in the presence of berberine (1.25, 2.5, 5 μM) were measured by q-RT-PCR and Western blot analysis. (E) Silencing of LH2 and validated the knockdown efficiency at protein levels. (F) Overexpression of LH2 and validated the efficiency at protein levels. (G–H) Representative images of the MDA-MB-231 or BT-549 cells treated with berberine (5 μM) or not after LH2 silencing. (I) Representative images of the MDA-MB-231cells treated with berberine (5 μM) or not after the LH2 overexpression. 10 × Scale bars, 200 μm. All results were representative of three independent experiments. **p < 0.01, ***p < 0.001 vs. control, nc, vehicle cells or LH2 overexpression cells. ###p < 0.001 vs. vehicle cells treated with berberine.